Separation and quantitation of the polyamine biosynthesis inhibitor D,L-alpha-difluoromethylarginine and other guanidine-containing compounds by high-performance liquid chromatography.
The arginine decarboxylase inhibitor difluoromethylarginine (DFMA) is an important tool in the study of polyamine metabolism, particularly with respect to the human pathogen Trypanosoma cruzi. This paper demonstrates a unique method for the detection and quantitation of intracellular DFMA using the fluorogenic agent 9,10-phenanthrenequinone. After separation of cell extracts by HPLC, DFMA can be accurately and reproducibly quantified with a lower sensitivity limit of 0.1 nmol by this simple fluorometric method. This assay can also be used to detect other guanidine-containing compounds such as arginine, agmatine, creatinine, and hirudonine, but not substituted guanidines such as aminoguanidine and creatine, or the structurally related amidines such as benzamidine and pentamidine.
Hunter
K J
KJ
Department of Medical Parasitology, London School of Hygiene and Tropical Medicine, United Kingdom.
Fairlamb
A H
AH
D000818:Animals; D001120:Arginine / Q000031:analogs & derivatives* / Q000032:analysis / Q000494:pharmacology; D002262:Carboxy-Lyases / Q000037:antagonists & inhibitors; D002851:Chromatography, High Pressure Liquid; D003421:Crithidia / Q000032:analysis* / Q000187:drug effects / Q000201:enzymology; D005456:Fluorescent Dyes; D019791:Guanidine; D006146:Guanidines / Q000032:analysis* / Q000494:pharmacology; D012680:Sensitivity and Specificity
D005456:Fluorescent Dyes; D006146:Guanidines; C030965:alpha-(difluoromethyl)arginine; D001120:Arginine; D002262:Carboxy-Lyases; C019722:arginine decarboxylase; D019791:Guanidine